• ISSN 1008-505X
  • CN 11-3996/S
杨丽娟, 王玉凤, 张翼飞, 付健, 于立红, 邓杰, 杨克军. 产酸克雷伯氏菌提高玉米幼苗耐盐碱胁迫的机理[J]. 植物营养与肥料学报, 2021, 27(6): 1044-1054. DOI: 10.11674/zwyf.2021011
引用本文: 杨丽娟, 王玉凤, 张翼飞, 付健, 于立红, 邓杰, 杨克军. 产酸克雷伯氏菌提高玉米幼苗耐盐碱胁迫的机理[J]. 植物营养与肥料学报, 2021, 27(6): 1044-1054. DOI: 10.11674/zwyf.2021011
YANG Li-juan, WANG Yu-feng, ZHANG Yi-fei, FU Jian, YU Li-hong, DENG Jie, YANG Ke-jun. Klebsiella oxytoca improves resistance of maize seedling to saline-alkali stress[J]. Journal of Plant Nutrition and Fertilizers, 2021, 27(6): 1044-1054. DOI: 10.11674/zwyf.2021011
Citation: YANG Li-juan, WANG Yu-feng, ZHANG Yi-fei, FU Jian, YU Li-hong, DENG Jie, YANG Ke-jun. Klebsiella oxytoca improves resistance of maize seedling to saline-alkali stress[J]. Journal of Plant Nutrition and Fertilizers, 2021, 27(6): 1044-1054. DOI: 10.11674/zwyf.2021011

产酸克雷伯氏菌提高玉米幼苗耐盐碱胁迫的机理

Klebsiella oxytoca improves resistance of maize seedling to saline-alkali stress

  • 摘要:
    目的 研究产酸克雷伯氏菌提高盐碱土壤生物学活性的效应,深入理解其缓解作物盐碱胁迫的机理,开发适用于盐碱土壤的微生物菌肥。
    方法 首先,诱导保存的产酸克雷伯氏菌产生可遗传的耐300 µg/L抗生素利福平的特性。然后,将产生抗性的菌种接种于LB液体培养基中,于30℃、180 r/min培养24 h,经离心、洗涤后用蒸馏水调配成1 × 108 cfu/mL的菌悬液,测定其固氮、溶磷、产铵和IAA能力,并逐步稀释制备1 × 106 cfu/mL、1 × 104 cfu/mL、1 × 102 cfu/mL菌悬液用于盆栽试验。盆栽试验用盐碱土 (pH 9.4) 采自黑龙江肇东市,供试作物为玉米,品种为郑单958。设置产酸克雷伯氏菌添加量为0、1 × 102、1 × 104、1 × 106、1 × 108 cfu/mL,施于土壤中,然后种植发芽的玉米。待玉米幼苗第二片叶展开后,测定其光合特性;幼苗生长12天后收获,调查玉米幼苗株高、根长、地上干重、地下干重,测定叶片中C、N和P含量。采用抖根法收集玉米幼苗根际土壤,测定土壤脲酶、碱性磷酸酶、蔗糖酶、过氧化氢酶活性。
    结果 产酸克雷伯氏菌发酵液的总固氮量为12.8 mg/L、可溶性磷含量85.59 mg/L、IAA浓度256.57 mg/L、产氨量75.46 mg/L。产酸克雷伯氏菌在盐碱土中可以很好地定殖,并在施入土壤后的第3天达到稳定的菌体数量。在盐碱土中,随着产酸克雷伯氏菌施用水平的增加,玉米幼苗根际土壤脲酶、碱性磷酸酶、蔗糖酶、过氧化氢酶活性总体呈现增加的趋势。与对照相比,施入产酸克雷伯氏菌水平为1 × 106、1 × 108 cfu/mL的处理4个酶活性增加幅度均达到显著水平,且以1 × 106 cfu/mL最适宜,相较CK脲酶活性提高62.81%、碱性磷酸酶活性提高17.74%、蔗糖酶活性提高115.5%、过氧化氢酶活性提高35.6%。与对照组相比,施用产酸克雷伯氏菌1 × 106 cfu/mL处理的植株叶片盐分离子含量显著减少,光合能力显著增强,提高玉米幼苗株高12.44%、根长22.59%、地上干重26.67%、地下干重37.78%。
    结论 产酸克雷伯氏菌具有固氮、溶磷和分泌生长素的功能。产酸克雷伯氏菌可显著提高根际土壤酶活性,降低玉米幼苗中Na含量,提高K和Ca2+含量以及K/Na和Ca2+/Na的值,进而显著提高幼苗的光合效率,促进玉米幼苗生长,提高玉米幼苗耐盐碱能力。但是,产酸克雷伯氏菌的促生效果取决于施用量,具体田间施用浓度还需进一步试验确定。

     

    Abstract:
    Objectives The effects of Klebsiella oxytoca on the biological properties of saline-alkali soil were studied to provide a theoretical basis for developing suitable microbial fertilizer in saline-alkali areas.
    Methods Zhengdan 958 variety of maize was used as the test material. Klebsiella oxytoca was induced to produce heritable resistant properties to 300 g/L antibiotic rifli. The resistant strains were inoculated in LB liquid medium and cultured at 180 r/min at 30℃ for 24 h. After centrifugation and washing, a bacterial suspension of 1×108 cfu/m was prepared using distilled water. Thereafter, its nitrogen fixation, phosphorus dissolution, ammonium production, and IAA capacity were measured. The bacterial suspensions of 1×106 cfu/mL, 1×104 cfu/mL, and 1×102 cfu/mL were gradually diluted and prepared for pot experiment. The saline-alkali soil (pH 9.4) was collected from Zhaodong City, Heilongjiang Province. Klebsiella oxytoca was added to the soil at 0, 1×102, 1×104, 1×106 and 1×108 cfu/mL, followed by the planting of germinated maize. The photosynthetic characteristics of maize seedlings were measured after the second leaf unfolds. The seedlings were harvested after 12 days of growth. Plant height, root length, aboveground dry weight, and underground dry weight of the maize seedlings were measured. The contents of C, N and P in maize leaves were determined. The activities of urease, alkaline phosphatase, sucrase and catalase in the rhizosphere of maize seedlings were determined using the root shaking method.
    Results The total nitrogen fixation, soluble phosphorus content, IAA concentration and ammonia yield of Klebsiella oxytoca were 12.8 mg/L, 85.59 mg/L, 256.57 mg/L and 75.46 mg/L , respectively. Klebsiella oxytoca could colonize well in saline-alkali soil and reached a stable number of bacteria on the third day after application. In saline-alkali soil, the activities of urease, alkaline phosphatase, acid phosphatase and catalase in rhizosphere soil of maize seedlings increased with the increase of Klebsiella oxytoca application level. Compared with the control, the four enzyme activity (urease activity, alkaline phosphatase activity, sucrase activity, and catalase activity) was significantly increased(P<0.05)in all the treatments, with 1×106 cfu/mL recording the highest value. The urease activity, alkaline phosphatase activity, sucrase activity, and catalase activity increased by 62.81%, 17.74%, 115.5%, and 35.6% compared with CK. Klebsiella oxytoca applied at 1×106 cfu/mL significantly reduced the salt ions in leaves, increased photosynthetic capacity, increased plant height by 12.44%, root length by 22.59%, aboveground dry weight by 26.67%, and underground dry weight by 37.78% compared with CK.
    Conclusions Klebsiella oxytoca is capable of fixing N, dissolving phosphorus and secreting auxin. Klebsiella oxytoca can significantly increase enzyme activity in the rhizosphere, decrease Na+ content in maize seedlings, increase the contents of K+ and Ca2+ and the ratio of K+/Na+ and Ca2+/Na+. Further, it significantly improves the photosynthetic efficiency and salt-alkali tolerance of maize seedlings and promotes the growth of maize. However, the effect of Klebsiella oxytoca depends on the application rate, and the specific field concentration requires further research.

     

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